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Figure 3: BA6 induced intracellular and mitochondrial ROS production and dissipation of MMP in A549 cells. (a) The probe of DCFH-DA (green) fluorescence was used to detect intracellular ROS (shifted towards the right) by flow cytometry in A549 cells treated without or with 10 μM BA6 for 24 h. (b) Relative quantification of BA6-induced cellular ROS production. Values are the of three independent experiments. (c) The probe of MitoSOX Red (red) fluorescence was used to detect mtROS (shifted towards the right) by flow cytometry in A549 cells treated without or with 10 μM BA6 for 24 h. (d) Relative quantification of BA6-induced mtROS production. Values are the of three independent experiments. (e) The mitochondrial membrane potential and depolarization were monitored by fluorescence dye of rhodamine 123 and JC-1 in A549 cells. Rhodamine 123 (green fluorescence) dyes were used to detect the membrane potential of the mitochondria. The histogram showed rhodamine 123 signals from untreated or treated 10 μM BA6 for 24 h, and the left shift represents the loss of MMP. (f) Relative quantification of BA6-induced disruption of MMP. The percentages of cells with intact MMP were decreased, along with increasing concentration of BA6. (g) The aggregate fluorescent count was indicative of mitochondrial depolarization by JC-1 staining and flow analysis assessment. JC-1 can be discriminated by a set gating measurement area. The presentative flow cytometry dot plots demonstrated A549 cells treated without or with 10 μM BA6 for 24 h. (h) Quantification analysis of the percentages of cells with depolarized mitochondria after treatment indicated concentrations of BA6 for 24 h. All total cells are 20,000 events, and the results are presented as the of three independent experiments. and vs. the untreated BA6 (0 μM) group.